Abstract：
In recent years, more sophisticated DNA technologies for genotyping have enabled considerable progress in various fields such as clinical genetics, archaeogenetics and forensic genetics. DNA samples previously rejected as too challenging to analyze due to low amounts of degraded DNA can now provide useful information. To increase the chances of success with the new methodologies, it is crucial to know the fragment size of the template DNA molecules, and whether the DNA in a sample is mostly single or double stranded. With this knowledge, an appropriate library preparation method can be chosen, and the DNA shearing parameters of the protocol can be adjusted to the DNA fragment size in the sample. In this study, we first developed and evaluated a user-friendly fluorometry-based protocol for estimation of DNA strandedness. We also evaluated different capillary electrophoresis methods for estimation of DNA fragmentation levels. Next, we applied the developed methodologies to a broad variety of DNA samples processed with different DNA extraction protocols. Our findings show that both the applied DNA extraction method and the sample type affect the DNA strandedness and fragmentation. The established protocols and the gained knowledge will be applicable for future sequencing-based high-density SNP genotyping in various fields.
摘要：
近年来,更复杂的DNA基因分型技术在临床遗传学等各个领域取得了相当大的进展，考古遗传学和法医遗传学。以前被拒绝的DNA样品由于低量的降解DNA而难以分析，现在可以提供有用的信息。为了增加新方法的成功机会，了解模板DNA分子的片段大小至关重要，以及样本中的DNA是单链还是双链。有了这些知识,可以选择合适的文库制备方法，和方案的DNA剪切参数可以根据样品中的DNA片段大小进行调整。在这项研究中,我们首先开发并评估了一种用户友好的基于荧光法的DNA片段估计方案.我们还评估了不同的毛细管电泳方法来估计DNA片段化水平。接下来,我们将开发的方法应用于使用不同DNA提取方案处理的各种DNA样品。我们的发现表明，所应用的DNA提取方法和样品类型都会影响DNA片段化和片段化。建立的方案和获得的知识将适用于未来的基于测序的高密度SNP基因分型在各个领域。
